IMMORTALIZED CEREBELLAR CELLS CAN BE INDUCED TO DISPLAY MATURE NEURONAL CHARACTERISTICS

We have established and characterized a cell line (designated Cb-E1A) that can be induced to display a variety of neuronal characteristics u nder simple culture conditions. This cell line was generated by retrov iral-mediated gene transfer of the adenovirus 12S E1A-immortalizing ge ne in cerebellar cells isolated from one-week-old rats. Actively divid ing cells express the E1A adenovirus protein, and exhibit minimal expr ession of glial cell markers and low level expression of neuronal cell markers. The immortalized cells can be induced to differentiate by cu lture in an alternative depolarizing medium or calcium ionophore-conta ining medium. This caused the expression of neuronal markers to increa se rapidly, while glial markers remain unchanged. Under these culture conditions, the Cb-E1A cells also display a variety of other character istics which suggest that they may provide a good model system for dif ferentiated cerebellar granule neurons. Such neuronal characteristics include a reduction or cessation of mitosis and an increased susceptib ility to glutamate toxicity. We think that this novel cell line and di fferentiation strategy will facilitate future studies of the cellular mechanisms involved in a wide variety of neuronal functions, including development and neurodegenerative disease. Copyright (C) 1996 IBRO. P ublished by Elsevier Science Ltd.